Gerald A. Denys

Levofloxacin Treatment Failure in a Patient with Fluoroquinolone-Resistant Streptococcus pneumoniae Pneumonia

The frequency of fluoroquinolone‐resistant Streptococcus pneumoniae has increased as fluoroquinolone administration for treatment of respiratory tract infections has increased. Levofloxacin treatment failed in a patient who had pneumococcal pneumonia and had received three previous courses of levofloxacin therapy. Susceptibility testing revealed high‐level resistance to levofloxacin (minimum inhibitory concentration [MIC] > 32 μg/ml), and cross‐resistance to moxifloxacin (MIC 4 μg/ml), trovafloxacin (6 μg/ml), and gatifloxacin (12 μg/ml). Sequencing of the quinolone‐resistance determining region revealed a mutation of serine‐81 to phenylalanine (Ser81→Phe) in the gyrA region of DNA gyrase and a Ser79→Phe mutation in the parC region of topoisomerase IV. The patient was treated successfully with intravenous ceftriaxone followed by oral cefprozil. Clinicians must be aware of local resistance patterns and the potential for fluoroquinolone treatment failures in patients with infections caused by S. pneumoniae.

Evaluation of BACTEC MYCO/F Lytic Medium for Recovery of Mycobacteria, Fungi, and Bacteria from Blood

ABSTRACT MYCO/F Lytic medium (MFL), a liquid medium developed for use with the BACTEC 9240 blood culture system, was compared to the Isolator system (IS) for the recovery of fungi and to the BACTEC 13A medium for the recovery of mycobacteria. Recovery of bacteria was compared to routine BACTEC Plus Aerobic/F (AF) blood cultures. Microbial growth was detected in 203 (17%) of 1,166 blood cultures. Fifty-seven specimens were positive for fungi: 35 were positive with both IS and MFL; six were positive with IS only (three Candida albicans, oneHistoplasma capsulatum, one Candida glabrata, and one Fusarium species isolate); three were positive with AF only (two C. albicans and one Candida parapsilosis isolate); and 13 were positive with MFL only (fiveC. glabrata, three C. albicans, twoCandida krusei, two Candida tropicalis, and oneC. parapsilosis isolate; P > 0.05 versus IS). Eighteen of 19 blood cultures positive for H. capsulatum grew in both IS and MFL, although the time to detection for MFL was greater. The mean time to detection for all fungi was 8.15 days for IS and 12.07 days for MFL. Seven hundred forty specimens were also cultured for mycobacteria with MFL and 13A. Forty-four grew mycobacteria; 38 were positive with both 13A and MFL; and 16 were positive with MFL only. Mycobacterium avium was recovered from 41 specimens; 36 were positive for both systems and 5 were positive for MFL alone. MFL was also compared to the AF bottle for the same 740 specimens. MFL and AF both detected 34 of the 40 clinically significant bacteria, while IS detected only 15 of 40. In summary, MFL is an excellent medium for the recovery of fungi, mycobacteria, and bacteria; however, the time to detection of H. capsulatum is increased.

Azithromycin treatment failure in community-acquired pneumonia caused by Streptococcus pneumoniae resistant to macrolides by a 23S rRNA mutation

In this report, we describe an azithromycin treatment failure in community-acquired pneumonia. During the first three days of azithromycin, the patients symptoms worsened, and she was subsequently admitted to the hospital. Blood cultures were positive for a penicillin-susceptible, macrolide-resistant S. pneumoniae. DNA sequencing revealed an A2059G mutation in domain V of the 23S rRNA. To our knowledge, this is the first clinical report of an azithromycin failure in the treatment of S. pneumoniae resistant to macrolides by this mechanism.

Multicenter Clinical Evaluation of the Portrait Toxigenic C. difficile Assay for Detection of Toxigenic Clostridium difficile Strains in Clinical Stool Specimens

ABSTRACT We compared the Portrait Toxigenic C. difficile Assay, a new semiautomated sample-to-result molecular test, to a toxigenic bacterial culture/cell cytotoxin neutralization assay (TBC/CCNA) for the detection of toxigenic Clostridium difficile in 549 stool specimens. Stool specimens were also tested by one of three alternative FDA-cleared molecular tests for toxigenic C. difficile (Xpert C. difficile, Illumigene C. difficile, or GeneOhm Cdiff). The sensitivities and specificities of the molecular tests compared to TBC/CCNA were as follows: 98.2% and 92.8% for the Portrait assay, 100% and 91.7% for the Xpert assay, 93.3% and 95.1% for the Illumigene assay, and 97.4% and 98.5% for the GeneOhm assay, respectively. The majority of Portrait false-positive results (20/31; 64.5%) were also positive for C. difficile by an alternative molecular test, suggesting an increased sensitivity compared to the culture-based “gold standard” method. The Portrait test detected an assay input of 30 CFU in 100% of spiked samples and detected an input of 10 CFU in 96.7% of samples tested.

Automated Detection of Toxigenic Clostridium difficile in Clinical Samples: Isothermal tcdB Amplification Coupled to Array-Based Detection

ABSTRACT Clostridium difficile can carry a genetically variable pathogenicity locus (PaLoc), which encodes clostridial toxins A and B. In hospitals and in the community at large, this organism is increasingly identified as a pathogen. To develop a diagnostic test that combines the strengths of immunoassays (cost) and DNA amplification assays (sensitivity/specificity), we targeted a genetically stable PaLoc region, amplifying tcdB sequences and detecting them by hybridization capture. The assay employs a hot-start isothermal method coupled to a multiplexed chip-based readout, creating a manual assay that detects toxigenic C. difficile with high sensitivity and specificity within 1 h. Assay automation on an electromechanical instrument produced an analytical sensitivity of 10 CFU (95% probability of detection) of C. difficile in fecal samples, along with discrimination against other enteric bacteria. To verify automated assay function, 130 patient samples were tested: 31/32 positive samples (97% sensitive; 95% confidence interval [CI], 82 to 99%) and 98/98 negative samples (100% specific; 95% CI, 95 to 100%) were scored correctly. Large-scale clinical studies are now planned to determine clinical sensitivity and specificity.

Fluoroquinolone susceptibility, resistance, and pharmacodynamics versus clinical isolates of Streptococcus pneumoniae from Indiana

The in vitro activity and pharmacodynamics (AUC(0-24)/MIC) of levofloxacin, gatifloxacin, moxifloxacin, and gemifloxacin were evaluated against 307 clinical isolates of Streptococcus pneumoniae from Indianapolis, Indiana. Organisms were collected between January 1999 and April 2000, and MICs were determined by broth microdilution. Serum concentration-time profiles were simulated for the following oral regimens administered once daily: levofloxacin 500 mg and 750 mg; gatifloxacin 400 mg; moxifloxacin 400 mg; gemifloxacin 320 mg. Free 24 h area under the serum concentration-time curves (AUC(0-24)) were calculated, and the average AUC(0-24)/MIC was calculated for each regimen. Differences in AUC(0-24)/MIC among agents were determined by analysis of variance (Scheffe post-hoc test, p < 0.05). Overall, gemifloxacin was the most potent agent tested. Five (1.7%), 4 (1.3%), and 2 (0.7%) isolates were resistant to levofloxacin, gatifloxacin, and moxifloxacin, respectively. None of the isolates was resistant to gemifloxacin. Gemifloxacin AUC(0-24)/MIC was significantly greater than all other regimens (p < 0.0001), with the exception of moxifloxacin. However, the percent of isolates for which an AUC(0-24)/MIC >or= 30-50 can be achieved is similar for gemifloxacin, moxifloxacin, gatifloxacin, and levofloxacin 750 mg. Large comparative studies are needed to determine if the differences in AUC(0-24)/MIC among fluoroquinolones are clinically significant.

Case report of Aurantimonas altamirensis bloodstream infection.

Luong et al. ([7][1]) recently described the first report and characterization of Aurantimonas altamirensis recovered from human clinical specimens. According to that report, the isolates were described as being “possibly associated” with infection. In April 2008, a 50-year-old male patient with

Serious streptococcal infections produced by isolates resistant to streptogramins (quinupristin/dalfopristin): case reports from the SENTRY antimicrobial surveillance program.

The emergence and sustained prevalence of Gram-positive organisms resistant to antimicrobials has been of interest for over a decade. Quinupristin/dalfopristin (formerly RP 59500 or Synercid) is a new injectable streptogramin combination that has been reported to have activity against Gram-positive organisms, even those with documented MLS(B) resistance. However, the two case reports presented here illustrate three well-documented Streptococcus spp. strains (S. mitis, S. pneumoniae) to be resistant to quinupristin/dalfopristin (MICs at 3, 8, and 12 microg/ml) following referral as routine isolates in the SENTRY Antimicrobial Surveillance Program. The S. pneumoniae pleural fluid isolate was cross-resistant to erythromycin. Both bacteremic S. mitis strains were resistant to macrolides (erythromycin, azithromycin, clarithromycin), lincosamides (clindamycin), and fluoroquinolones. Patient histories indicated no prior use of MLS class antimicrobials for the S. mitis case, but the patient having the S. pneumoniae isolate did receive prior treatment of erythromycin and clindamycin. All isolates had modestly increased penicillin MICs of 0.12 microg/ml. The mode of resistance to quinupristin/dalfopristin was not evident (sat A-negative by PCR); and these cases illustrate the existence of streptogramin-resistant isolates before the introduction of this antimicrobial class into human clinical practice.

Propionibacterium acnes, Coagulase-Negative Staphylococcus, and the "Biofilm-like" Intervertebral Disc.

Study Design. Patients scheduled for spinal surgery were screened prospectively for a microbial presence associated with intervertebral disc specimens. Inclusion was limited to patients requiring surgery for any of five conditions: study patients with cervical spine intervertebral herniation (IVH), lumbar spine IVH, lumbar spine discogenic pain, and control patients with idiopathic scoliosis/Scheurermanns kyphosis or trauma/neuromuscular deformity. Exclusion criteria included ongoing systemic infection, abnormal pre-operative white cell counts, documented or suspected spinal infection, or previous surgery to the involved disc. Objective. The aim of this study was to test for an association between the presence of a bacterial entity in operated discs and a diagnosis of pathologic disc disease. Summary of Background Data. An association has been described between microbial colonization and progressive intervertebral disc degeneration in 36 herniation patients undergoing microdiscectomies. A total of 19 patients had positive cultures on long-term incubation, with Propionibacterium acnes present in 84% of discs. Materials and Methods. Discs were harvested during surgery, using strict sterile technique. Each disc was divided, with half the sample sealed in a sterile, commercially prepared anaerobic culture transport container, and half fixed in formalin. Live specimens were cultured for bacteria at a university-affiliated laboratory in a blinded fashion. Fixed pathologic specimens were gram-stained and read by a board-certified pathologist. Results. A total of 169 intervertebral discs from 87 patients were evaluated (46 males, 41 females). Positive cultures were noted in 76 of 169 discs (45%), with 34 discs positive for P. acnes and 30 discs positive for Staphylococcus. No pathologic evidence was seen of microorganisms, acute or chronic inflammation, or infection. Pooling the IVH and discogenic pain patients and contrasting them with control patients showed a significant association of IVH with positive bacterial cultures (&khgr;2 = 15.37; P = 0.000088). Conclusion. Endemic bacterial biofilms are significantly associated with IVH and discogenic pain. Level of Evidence: N/A

Selection of a gyrA mutation and treatment failure with gatifloxacin in a patient with Streptococcus pneumoniae with a preexisting parC mutation.

An 81‐year‐old woman had pneumonia caused by Streptococcus pneumoniae (levofloxacin Etest minimum inhibitory concentration [MIC] 1.5 μg/ml) and was treated with intravenous gatifloxacin 200 mg/day. After 3 days of therapy, repeat sputum cultures were positive for S. pneumoniae, which was resistant to levofloxacin (Etest MIC > 32 μg/ml). The isolate obtained before therapy showed a preexisting parC mutation of aspartic acid‐83 to asparagine (Asp83→Asn), and the isolate obtained during therapy showed an acquired gyrA mutation from serine‐81 to phenylalanine (Ser81→Phe) and a second parC mutation from lysine‐137 to Asn (Lys137→Asn). Both isolates were the same strain, as determined with pulsed‐field gel electrophoresis. This case demonstrates the potential for resistance to emerge during 8‐methoxy fluoroquinolone therapy for fluoroquinolone‐susceptible S. pneumoniae with a preexisting parC mutation. Additional clinical failures with a fluoroquinolone may occur unless these first‐step parC mutants can be identified to assist clinicians in selecting appropriate antimicrobial therapy.